Gene expression profiling of substantia nigra: a post-mortem study in Parkinson's disease and control subjects
Gene expression profiling of substantia nigra: a post-mortem study in Parkinson's disease and control subjects
Human midbrain samples were provided by the Parkinson's UK Brain Bank (Imperial College London, London, England).  RNA was extracted starting from 5 µm FFPE slide-mounted sections using RecoverAll Total Nucleic Acid Isolation Protocol (ThermoFischer Scientific), according to the manufacturer's instructions.  The RNA was stored at -80° C until further processing. RNA sequencing and data analysis were performed by Genomix4Life Srl (Baronissi, Italy). The yield and quality of RNA were evaluated by  Qubit fluorometer (Thermo Fisher Scientific), and TapeStation 4200 (Agilent Technologies 5301 Stevens Creek Blvd Santa Clara, CA USA), respectively. Indexed libraries were prepared from 50ng/ea purified RNA with Illumina Stranded Total RNA with Ribo-Zero Plus (Illumina) Kit according to the manufacturer's instructions. For library quantifications, TapeStation 4200 (Agilent Technologies) and Qubit (Thermo Fischer) was used. Indexed libraries were pooled in equimolar amounts with a final concentration of 2 nM.